Biomedical subjects
Publications and source records attributed to R Klieber.
The exchange interaction for the yellow 1S orthoexciton in Cu2O is derived up to the order K2. The resulting exchange splittings are verified experimentally using high resolution spectroscopy. In agreement with theory the fine structure shows a characteristic dependence on the direction of the wave vector.
Immunochemical studies were undertaken to identify surface-orientated epitopes of the free alpha subunit of human chorionic gonadotrophin (hCG-alpha) at the amino acid sequence level. We investigated the molecular organization of these epitopes, resolved the immunological topography in terms of spatial arrangement of antigenic domains and related structures to functions such as subunit association or receptor binding. Overlapping synthetic peptides covering the entire amino acid sequence of hCG-alpha, an enzymatically digested hCG-alpha subunit, and a reduced and alkylated hCG-alpha preparation were assayed in a solid-phase one-site enzyme-linked immunoassay, and in a solution-phase competitive radioimmunoassay (RIA). The antigenic topography was mapped by monoclonal antibodies (MCAs) in two-site binding assays (sandwich RIA). On the surface of hCG-alpha, seven different epitopes (alpha 1-alpha 7), arranged in four spatially distinct domains, could be distinguished: A, alpha 1,2,4; B, alpha 3,5; C, alpha 6; D, alpha 7. The peptides spanning hCG-alpha(13-18), hCG-alpha(17-22) and hCG-alpha(33-42) appeared to contribute to the formation of epitopes alpha 2, alpha 4 and alpha 6 respectively. Since epitope alpha 6 is present only on the free non-assembled subunit of different species, we concluded that the region hCG-alpha(33-42), which is evolutionarily highly conserved, represents a subunit assembly site. All but one epitope (alpha 7) are destroyed by reducing and alkylating hCG-alpha. In contrast, chymotryptic digestion of hCG-alpha, leading to release of the heptapeptide hCG-alpha(41-47), did not affect epitope expression, indicating that this sequence is not involved in the formation of antigenic determinants. Addressing the biological properties of hCG-alpha epitopes by radioreceptor assay revealed that the three hCG-alpha peptides corresponding to epitopes alpha 2, alpha 4 and alpha 6 did not displace radiolabelled hCG from its receptor, whereas any of the MCAs directed against determinants (alpha 1-alpha 5), shared by hCG and hCG-alpha, totally inhibited binding. Consistent with this, the antibodies neutralized the biological activity of hCG in terms of testosterone production in a mouse Leydig cell in vitro bioassay. We therefore concluded that hormone antibody-binding sites differ from those of hormone receptor binding, revealing no essential congruence of immunologically and biologically active domains.
The World Health Organisation (WHO) Task Force on Birth Control Vaccines has selected the pregnancy hormone human chorionic gonadotropin (hCG) as a target molecule for a contraceptive vaccine. A synthetic peptide antigen corresponding to the amino acid sequence 109-145 of the carboxyl-terminal portion of the hCG beta-subunit (hCG beta CTP), which is supposed to elicit hCG-immunoneutralizing antibodies, has been submitted to clinical trials. Recent findings suggest that hCG beta CTP does not play a role in the biological activity of hCG. This raises the question concerning the assumed mechanism of action of the hCG beta CTP-based birth control vaccine. We therefore investigated the immunoneutralizing capacity of antibodies directed against hCG beta CTP. Although it is possible to generate specific monoclonal and polyclonal antibodies for hCG by using hCG beta CTP as an immunogen, it appeared that the biological response to hCG was not affected by such antibodies. The reason for this is that the hCG-antibody-complex is still able to bind to target cell receptors and therefore the intended contraceptive effect should not occur. In addition there is a risk of hazardous possible side effects such as an autoimmune reaction against the ovary because we found that at least one epitope is still accessible for antibody binding on receptor-bound hCG. We conclude from our results that both the efficacy and safety of the WHO vaccine are not yet ensured.
To evaluate in vivo the proposed intrinsic thyroid-stimulating hormone (TSH) activity (TSA) of human chorionic gonadotropin (hCG), we monitored over 0.5-1 years the thyroid status of eight patients with hCG-producing non-seminomatous testicular cancer. The patients' sera were analyzed for concentrations of hCG, free thyroxine (fT4), hTSH, and thyroxine-binding globulin (TBG). All patients had excessively high concentrations of hCG (1 x 10(5)-5 x 10(8) ng/L, mean: 1 x 10(7) ng/L) before polychemotherapy, which decreased under successful therapy to physiological values (< 240 ng/L). Although the serum concentrations of hCG varied by more than six orders of magnitude, we saw no changes and no correlation (P > 0.05) between the concentrations of hCG and the concentrations of fT4 and hTSH. Not even when hCG concentrations were greatest (> 5 x 10(7) ng/L) were any signs of hyperthyroidism observed: fT4 (3.5-13 ng/L) and hTSH (9-700 ng/L) were in the physiological range in all patients and remained so during chemotherapy. The results of this longitudinal study were confirmed in analyzing the data for all eight patients (total: 82 samples) cross-sectionally. Again, we found no correlation (P > 0.05) between the concentrations of hCG and fT4 or hCG and hTSH. We conclude that even excessive amounts of testicular tumor-derived hCG do not display any TSH-like activity in vivo.
To determine the serum concentrations of human chorionic gonadotropin (hCG), its free beta-subunit (hCG beta), and the free alpha-subunit (free alpha) common to all human glycoprotein hormones under physiological and pathological conditions, we developed monoclonal antibody-based immunoenzymometric assays. Free alpha-subunit was detected in the sera of all healthy individuals of both sexes; hCG was measurable in sera of 54% of the men, and 46% were positive for free hCG beta; in nonpregnant women, 69.5% were positive for hCG, 68.4% for the free beta-subunit. Pathological conditions, i.e., hCG-producing tumors, were studied in vitro and in vivo. In vitro, the concentrations of hCG, free hCG beta, and free alpha in tissue-culture supernates of a choriocarcinoma cell-line ("JAR") showed a parallel pattern during time-course analysis. In vivo, in long-term follow-up studies of 13 patients with testicular cancer, serum concentrations of the three analytes paralleled each other, whether the disease was in remission or not. Because of a selective increase of free hCG beta and free alpha in 27% of seminomatous tumor patients and in 13% of the nonseminomatous patients, the percentage of tumor-marker-positive sera was increased from 15% to 42% and 57% to 70%, respectively, by the additional measurement of free hCG beta and free alpha. Thus hCG, free hCG beta, and free alpha are physiologically present in a high percentage of the sera from healthy men, and the determination of free hCG beta and free alpha, although not of prognostic value, improves the diagnostic possibilities in patients with testicular cancer.
A previously established map of the surface epitopes of human chorionic gonadotropin (hCG) served as template for the present study in which we investigated the antigenic surfaces of two glycosylation variants of hCG, i.e. desialylated hCG (asialo-hCG) and deglycosylated hCG (degly-hCG). This map allocates five epitopes to the alpha subunit, five to the beta subunit and four alpha beta epitopes to structures formed only by the alpha/beta heterodimer holo-hCG (Schwarz et al. (1986) Endocrinology 118, 189-197; Berger et al. (1990) J. Endocrinol. 125, 301-309). Here it is described that both variants complied with this template: each of the 14 distinct monoclonal antibodies with which the epitopes of hCG were defined reacted with radiolabeled asialo-hCG and degly-hCG as well and generally bound degly-hCG with greater affinity than hCG. Moreover, every combination of capture and radiolabeled detection antibody that was either compatible or incompatible on unlabeled hCG was so also on unlabeled asialo-hCG and degly-hCG. It thus appears that alterations of the carbohydrate structure of hCG can be associated with a change in affinity between some antibodies and their respective epitopes but not with a loss of an epitope or with a change in the topographical relationships of the 14 epitopes.
Immunization with purified cardiac myosin induces autoimmune myocarditis in A strain mice. Because this disease parallels Coxsackievirus B3 (CB3)-induced myocarditis in many respects, we are now using the immunization model as a virus-free system to study certain forms of immunologically-mediated heart disease. In the present article we will describe several characteristics of the humoral and cellular autoimmune response acting in myosin-induced myocarditis. Furthermore, we discuss hypotheses which might explain how myosin-reactive T cells recognize and attack the heart tissue.
Explore the source record for details and available documents.
Using a panel of monoclonal antibodies (MCA) to human chorionic gonadotropin (hCG) and its alpha and beta subunits and additional MCA to the non-assembled, free alpha and beta chains which were produced in the course of the present experiments, it was possible to extend the previously established epitope map of hCG. Two MCAs turned out to be specific for the free alpha chain of the glycoprotein hormones (GPH) and, thus, did not react with holo hCG. Two other MCA recognized two epitopes (beta 6 and beta 7) on the free form of the hCG beta only. Again, no holo hormone cross-reaction was observed. Whereas previously only nine antigenic hCG determinants (3 alpha, 4 beta, 2c) had been demonstrated, it was now possible to distinguish fourteen epitopes (5 alpha, 5 beta, 4c). In addition, epitope maps were established for the non-assembled, free subunits of hCG. These comprise six epitopes on the alpha chain (alpha 1- alpha 6) and seven on the beta chain (beta 1- beta 7). Both so far generally accepted premises of Judith Vaitukaitis, claiming the beta chain of the GPH to be immunologically species-specific, the beta chain to be immunologically hormone-specific, were clearly disproven by demonstrating inter- and intra-species cross-reaction of some MCA. Based on the immunological topography of the holo hCG molecule and its free subunits, highly specific and sensitive immuno-enzymometric assays (IEMA) with predictable specificities were designed, measuring either non-assembled subunits alone or in combination with the intact hormones.
Discordant results on body fluid levels of human chorionic gonadotrophin (hCG) free alpha- and beta-subunits under physiological and pathophysiological conditions, prompted us to raise a total of 260 monoclonal antibodies (MCA) against free hCG-alpha, free hCG-beta, holo-hCG, human follicle-stimulating hormone and bovine luteinizing hormone; 153 MCA recognizing the human alpha-subunit and 28 reacting with hCG-beta were extensively analysed for their intra- and inter-species cross-reactivity with homologous hormones, and for the compatibility of epitopes recognized by them. The immunological topography of free hCG-alpha and free hCG-beta was resolved by these MCA, and epitope maps were designed. Six antigenic determinants on the free alpha-chain (alpha 1-alpha 6), clustered in three spatially distinct domains, and seven epitopes on the surface of free hCG-beta (beta 1-beta 7), could be distinguished. Strikingly, three alpha-chain epitopes (alpha 4, alpha 5 and alpha 6) were shared between various species, which is in contradiction to the concept of immunological species-specificity of alpha-subunits. Three determinants were found to be present only on the free subunits but not on holo-hCG (alpha 6, beta 6 and beta 7), and only two determinants (beta 1 and beta 7) were hormone-specific for hCG. Based on this information, an immunoenzymometric assay for the free alpha-subunit of human glycoprotein hormones was established, with a sensitivity of 1.3 pg/well and a cross-reactivity with holo-hCG of less than 0.005%.(ABSTRACT TRUNCATED AT 250 WORDS)
Having recently analyzed with monoclonal antibodies (MCA) the immunologic surface of human chorionic gonadotropin (hCG) as consisting of 9 distinct epitopes exposed on the molecule in a characteristic topographical manner (Schwarz, S., Berger, P., and Wick, G., Endocrinology 118, 189-197, 1986) we now attempted to confirm this result on a more general basis, e.g. by incorporating MCA that were just recently obtained and previously not included. What we were, however, interested most was the question whether the tentative model of the epitope map of hCG could represent a "target" with which hCG-related hormones such as LH, FSH, and TSH (the family of glycoprotein hormones, GPH) would (partially) match. Indeed, repeating various immunizations with GPH of human as well as animal origin revealed a remarkable reproducibility in terms of several anticipated epitope specificities of MCA. This indicates that MCA can be regarded as reliable probes for mapping epitopes and, as we have presumed, of receptor interaction domains of GPH as well. Extending the originally used 2-site MCA binding exclusion approach by an interspecies crossreactivity (Xr) analysis we now are able to refine our epitope model of hCG such that 2 additional epitopes were found which were not previously resolvable. Most surprisingly, two of 5 epitopes on the alpha subunit were now also detected on various non-human GPH, which is in striking contrast to a seemingly well established dogma. Yet all five alpha epitopes of hCG are present on hLH, hFSH, and hTSH as well and arranged in the same spatial relationship to each other as on hCG. Even the 2 conformational epitopes and their close topographical relationship to the alpha-epitopes appear to be remarkably conserved on all human GPH. Among the beta-epitopes we have found one that is not shared by hLH and that - surprisingly - is not the C-terminal peptide (CTP) by which hLH differs from hCG. On the basis of this refined epitope map a way was paved along which it should be feasible to elucidate the sterical relationship of the epitopes to the receptor interaction domain(s) of hCG. To this end the MCA were tested in principally two ways: first, as to which of the 11 MCA with different epitope specificities would be able (or not) to inhibit by preincubation the binding of radiolabeled hCG (or hLH, respectively) to rat testis LH/hCG receptors? Secondly (and inversely) which of the 11 epitopes of hCG would still be accessible to binding by radiolabeled MCA when the (unlabeled) hormone is bound to the receptor?(ABSTRACT TRUNCATED AT 400 WORDS)